Inhibition of Ceramide Metabolism Sensitizes Human Leukemia Cells

Acetaminophen (paracetamol N-acetyl-p-aminophenol [APAP]) is a widely used analgesic/antipyretic drug with

Posted on March 9, 2016 at 5:12 am.

Acetaminophen (paracetamol N-acetyl-p-aminophenol [APAP]) is a widely used analgesic/antipyretic drug with few side effects at therapeutic doses [1]. as cytokeratin 18 [7] and high mobility group box 1 protein [8] are also observed in serum of patients with APAP-induced liver injury. Hepatocellular necrosis is initiated by a reactive metabolite N-acetyl-p-benzoquinone imine (NAPQI) mainly produced by cytochrome P450 (CYP) 2E1 [9-12]. NAPQI likely mediates injury via hepatic glutathione depletion oxidative and nitrosative stress and inflammation. Furthermore the development and progression of liver injury induced by APAP appears to involve multiple mediators including reactive oxygen species [13] peroxynitrite [14 15 cytokines [16-19] and eicosanoids [20 IL20 antibody 21 buy 529-44-2 Eicosanoids such as prostaglandins (PGs) and thromboxanes play an important role in the development of various diseases as well as APAP hepatotoxicity [22-25]. North et al. [21] and Cavar et al. [26] exhibited that PGE2 has a protective function in APAP-induced liver organ damage in mice and zebrafish respectively. PGI2 (prostacyclin) also appears to become a hepatoprotectant in APAP-induced liver organ injury [27]. Furthermore Reilly et al. [28] recommended that cyclooxygenase (COX) 2-produced PGs such as for example PGD2 and PGE2 possess a hepatoprotective function in APAP-induced liver organ damage in mice. On the other hand thromboxane A2 (TXA2) appears to exacerbate APAP hepatotoxicity. Ketoconazole buy 529-44-2 OKY-1581 and benzyl imidazole which inhibit TXA2 creation have the ability to prevent liver organ damage induced by APAP in rodents [20 29 These observations claim that inhibiting TXA2 creation is a appealing strategy for the treating liver organ injury because of APAP overdose. Nevertheless ketoconazole originally an antifungal agent provides side effects due to inhibition of CYPs [30]. Furthermore while OKY-1581 and benzyl imidazole are generally found in the lab they aren’t approved for scientific use. Which means potential customer for the scientific usage of TXA2 synthase inhibitors for the treating APAP-induced liver organ injury is normally unclear. Ozagrel (OKY-046; (E)-3-[4-(imidazol-1-ylmethyl)phenyl]prop-2-enoic acidity) originated being a selective TXA2 synthase inhibitor and continues to be trusted for treating sufferers with bronchial asthma and cerebral thrombosis and vasospasm in Japan [22 31 32 Ozagrel alleviates the symptoms of varied diseases as evaluated by biochemical and scientific evaluation [22 23 31 Nevertheless little continues to be reported on the consequences of ozagrel on APAP hepatotoxicity. Within this research we analyzed whether ozagrel could drive back APAP-induced liver organ damage in mice. We examined the effects of ozagrel on serum alanine aminotransferase (ALT) levels mortality and histological changes induced by APAP treatment. In addition we assessed hepatic glutathione content material and the manifestation of cell death-related markers in the APAP treated mice as well as CYP2E1 activity in mouse liver microsomes. Furthermore we investigated the effects of ozagrel on NAPQI-induced hepatic injury in vitro. Methods Reagents Ozagrel hydrochloride monohydrate was kindly donated by Ono buy 529-44-2 Pharmaceutical CO. LTD. (Osaka Japan) and Kissei CO. Ltd. (Nagano Japan). APAP NAPQI and NAC were purchased from Sigma-Aldrich (St. Louis Missouri USA). Metaphosphoric acid was purchased from Alfa Aesar (Ward Hill Massachusetts USA). Cell tradition reagents were from Gibco?-Existence Technologies (Existence Systems Japan Tokyo Japan). HyClone? fetal bovine serum (FBS) was purchased from Thermo Scientific (Logan UT USA). The cell counting kit and Cellstain? Double Staining Kit were from Dojindo Laboratories (Kumamoto Japan). All other reagents and solvents were of reagent grade. De-ionized and distilled bio-pure grade water was used throughout the study. Animal experiments The APAP overdose-induced liver injury model was based on a model previously reported by our group [34]. Male 7-9-week-old ICR mice (Charles River Laboratories Japan INC. Yokohama Japan) were used. Animals were housed in cages in a room under controlled conditions at 24°C having a 12-h light cycle and given free access to food and water. Mice were buy 529-44-2 fasted over night but given.